Project description:Offspring health outcomes are often linked with epigenetic alterations triggered by maternal nutrition and intrauterine environment. Strong experimental data also link paternal preconception nutrition with pathophysiology in the offspring, but the mechanism(s) routing effects of paternal exposures remain elusive. Animal experimental models have highlighted small non-coding RNAs (sncRNAs) as potential regulators of these effects. This study characterised the baseline sncRNA landscape of human sperm and the effect of a 6 week dietary intervention on their expression profile. 5’tRFs, miRNA and piRNAs were the most abundant sncRNA subtypes identified; their expression was associated with age, BMI and sperm quality. Nutritional intervention with vitamin D and omega-3 fatty acids altered expression of 3 tRFs, 15 miRNAs and 112 piRNAs, targeting genes involved in fatty acid metabolism and transposable elements in the sperm genome.
Project description:DNA from expanded bovine blastocysts and bovine sperm were extracted then subjected to methylation-sensitive enzymatic digestion and LM-PCR enrichment before being hybridized onto a microarray. Two-condition experiment, bovine blastocysts (pools of 10) vs bovine sperm. Four biological replicates of each tissue were hybridized to four two-color arrays in a dye-balanced design.
Project description:Identification of bovine sperm surface carbohydrate-binding proteins relevant in two important aspects of fertilization: (i) formation of the sperm reservoir in the oviductal epithelium, and (ii) gamete recognition (oocyte-sperm interaction). Using whole sperm cells and a novel affinity capture method that combines proteolysis of protein-glycan complexes and mass spectrometry (i.e., CREDEX-MS), several lectins were enriched, identified by MS/MS proteomics, and mapped within the fertilization events in bovine species.
Project description:DNA from expanded bovine blastocysts and bovine sperm were extracted then subjected to methylation-sensitive enzymatic digestion and LM-PCR enrichment before being hybridized onto a microarray.
Project description:Purpose: to construct a library of miRNAs in bovine sperm was constructed using Illumina high‐throughput sequencing technology. Result: Unique sequences that were 18–26 nucleotides in length were mapped to specific precursors in miRBase 20.0 using BLAST. A total of 951 known miRNAs and 8 novel, highly expressed miRNA candidates were identified.
Project description:Paternal exposure to a range of environmental and lifestyle factors elicits distinct changes to the sperm sncRNA profile; modifications that have significant post-fertilization consequences. Despite this knowledge, there remains limited mechanistic understanding of how paternal exposures effect the sperm sncRNA landscape. Here, we report the acute sensitivity of the sperm sncRNA profile to the potent reproductive toxicant, acrylamide. Further, we traced the differential accumulation of acrylamide responsive sncRNAs to coincide with sperm transit of the proximal (caput) segment of the epididymis, wherein acrylamide exposure altered the expression of several transcription factors implicated in the expression of acrylamide-sensitive sncRNAs. We also identified extracellular vesicles secreted from the caput epithelium in relaying altered sncRNA profiles to maturing spermatozoa, the implications of which manifest in the form of dysregulated gene expression during early embryonic development. These data provide a causative mechanistic link to account for how environmental insults can alter the sperm epigenome and compromise the transcriptomic profile of early embryos
Project description:Mammalian fertilization requires a complex interplay between a sperm cell and an oocyte. A significant proportion of these functions are mediated through the proteomic composition of the sperm surface. These proteomic constituents can present as biomarkers to control and regulate breeding of agricultural animals. Previous studies have addressed the composition of bovine sperm cell apical plasma membrane proteome with nitrogen cavitation enrichment. Alternative workflows would enable to expand the compositional data more globally around the entire sperm’s surface. We used a cell-surface biotin-labelling in combination with differential centrifugation to enrich bovine sperm surface proteins. Using nano-LC MS/MS, 338 proteins were confidently identified in the plasma membrane enriched proteome. These results were bioinformatically analysed to assess the purification and to highlight the potential protein-protein interaction networks on the sperm cell surface. A highly significant degree of enrichment was found for transmembrane and plasma membrane proteins, among them proteins previously not described on bovine sperm (CPQ, CD58, CKLF, CPLV, GLB1L3 and LPCAT2B). Functional categories of sperm-egg interaction, protein turnover, metabolism as well as molecular transport, spermatogenesis and signal transduction were represented by proteins with high quantitative signal in our study. A descriptive overview of the bovine sperm plasma membrane integral and peripheral proteins is provided to complement future studies on animal reproduction and its relation to sperm cell surface.
Project description:Paternal exposure to environmental stressors elicits distinct changes to the sperm sncRNA profile; modifications that have significant post-fertilization consequences. Despite this knowledge, there remains limited mechanistic understanding of how paternal exposures modify the sperm sncRNA landscape. Here, we report the acute sensitivity of the sperm sncRNA profile to the reproductive toxicant, acrylamide. Further, we traced the differential accumulation of acrylamide-responsive sncRNAs to coincide with sperm transit of the proximal (caput) segment of the epididymis, wherein acrylamide exposure altered the abundance of several transcription factors implicated in the expression of acrylamide-sensitive sncRNAs. We also identified extracellular vesicles secreted from the caput epithelium in relaying altered sncRNA profiles to maturing spermatozoa, and dysregulated gene expression during early embryonic development following fertilisation by acrylamide-exposed spermatozoa. These data provide mechanistic links to account for how environmental insults can alter the sperm epigenome and compromise the transcriptomic profile of early embryos.